Can We Use Entomopathogenic Fungi As Endophytes for Dual Biological Control of Insect Pests and Plant Pathogens? ⇑ Lara R

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Biological Control xxx (2017) xxx–xxx

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Biological Control

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Review Can we use entomopathogenic fungi as endophytes for dual biological control of insect pests and plant pathogens? ⇑ Lara R. Jaber a, , Bonnie H. Ownley b a Department of Plant Protection, Faculty of Agriculture, The University of Jordan, 11942 Amman, Jordan b Department of Entomology and Plant Pathology, The University of Tennessee, Knoxville, TN 37996-4560, USA highlights

Fungal entomopathogens as endophytes are garnering increased research attention. These agents have a promising potential for biocontrol of insect and pathogen pests. Consistent plant colonization must be established to achieve endophytic protection. Interaction with other endophytes should be considered to attain optimal efﬁcacy. Elucidating modes of action is essential to realize their full biocontrol potential. article info abstract

Article history: An increasing number of recent studies demonstrate that entomopathogenic fungi, often solely consid- Received 2 December 2016 ered as insect pathogens, play additional roles in nature, including endophytism, plant disease antago- Revised 21 January 2017 nism, plant growth promotion, and rhizosphere colonization. These newly emerging, but not yet fully Accepted 23 January 2017 understood, ecological roles hint at the possibility that we have been overlooking important attributes Available online xxxx in our quest to develop fungal entomopathogens exclusively as inundative biopesticides against insect and other arthropod pests. Such additional roles recently-discovered to be played by entomopathogenic Keywords: fungi provide opportunities for the multiple use of these fungi in integrated pest management (IPM) Dual microbial control agents strategies. Of particular interest is the ability displayed by various genera of entomopathogenic fungi Endophytes Fungal entomopathogens to colonize a wide variety of plant species in different families, both naturally and artiﬁcially following Hypocreales inoculation, and confer protection against not only insect pests but also plant pathogens. This article Insect pests reviews the literature currently available on the endophytic colonization of different host plants by fun- Integrated pest management (IPM) gal entomopathogens, and summarizes the negative effects of such colonization on insect pests and plant Mode of action pathogens that have been reported to date. It also addresses the possible mechanisms of protection con- Plant diseases ferred by endophytic fungal entomopathogens and explores the potential use of these fungi as dual microbial control agents against both insect and pathogen pests. Moreover, interactions amongst endophytic fungal entomopathogens and other endophytes are discussed. Finally, current limitations and future research directions for the innovative use of endophytic fungal entomopathogens as dual microbial control agents are summarized. Ó 2017 Elsevier Inc. All rights reserved.

Contents

1. Introduction ...... 00 2. Fungal entomopathogens as endophytes ...... 00 3. Endophytic fungal entomopathogens and potential for biological control of insect pests ...... 00 4. Endophytic fungal entomopathogens and potential for biological control of plant diseases ...... 00 5. Possible mechanisms of plant disease control by endophytic fungal entomopathogens...... 00

DOI of original article: http://dx.doi.org/10.1016/j.biocontrol.2017.01.013 ⇑ Corresponding author. E-mail address: [email protected] (L.R. Jaber). http://dx.doi.org/10.1016/j.biocontrol.2017.01.018 1049-9644/Ó 2017 Elsevier Inc. All rights reserved.

5.1. Direct suppression of plant pathogens...... 00 5.2. Induction of systemic plant resistance...... 00 5.3. Promotion of plant growth ...... 00 6. Interaction of endophytic fungal entomopathogens with other endophytes ...... 00 7. Conclusions and future prospects ...... 00 Acknowledgments ...... 00 References ...... 00

1. Introduction Even though many other definitions have been used ever since (see Box 1 in Hyde and Soytong, 2008), one common denominator Fungal entomopathogens, often solely considered as insect amongst them is that endophytes are organisms (fungi or bacteria) pathogens, have been studied for over a hundred years without that colonize internal plant tissues without causing apparent reaching their full potential as effective microbial control agents; symptoms or harm to their plant host. So defined, endophytes especially under field conditions (Vega et al., 2009). A growing comprise a diverse polyphyletic group of microorganisms that body of literature hints at the possibility that we have been over- can exhibit more than one type of life history at distinct life stages looking important attributes of fungal entomopathogens in our (Arnold and Lewis, 2005). quest to develop these microorganisms exclusively as biopesti- Although ubiquitous amongst terrestrial plants (Petrini, 1991; cides. An increasing number of recent studies demonstrate that Arnold et al., 2001), the majority of endophyte research has entomopathogenic fungi play additional roles in nature, including focused to date on the vertically-transmitted endophytes within endophytism, plant disease antagonism, plant growth promotion, the genus Neotyphodium (Clavicipitaceae) that systemically colo- and rhizosphere colonization. These newly emerging, but not yet nize the above-ground parts of some grasses. These clavicipita- fully understood, ecological roles provide opportunities for the ceous endophytes are generally known to confer an array of multiple use of fungal entomopathogens in integrated pest man- potential fitness benefits to their grass host plants (Kuldau and agement (IPM) strategies (Vega et al., 2009; Lacey et al., 2015). Bacon, 2008). Less attention has been given to the horizontally- Various genera of entomopathogenic fungi have recently been transmitted non-clavicipitaceous endophytes, which are wide- shown to act as plant endophytes in a variety of host plants spread in nature and dominated by the Ascomycetes (Arnold and (reviewed in Section 2 below), antagonists to plant pathogens Lutzoni, 2007); of which several genera are fungal ento- (Goettel et al., 2008; Ownley et al., 2008, 2010; Sasan and mopathogens (Ascomycota: Hypocreales). Bidochka, 2013; Jaber and Salem, 2014; Jaber, 2015), plant growth Emerging as an exciting new area of research, ‘fungal ento- promoters (Kabaluk and Ericsson, 2007; Garcia et al., 2011; Sasan mopathogens as endophytes’ has only rather recently been incor- and Bidochka, 2012; Liao et al., 2014; Lopez and Sword, 2015; porated into an over 100-year-old endophyte research base Jaber and Enkerli, 2016, 2017), and beneficial rhizosphere colonizers following the recovery of various genera of fungal ento- (Hu and St. Leger, 2002; St. Leger, 2008; Bruck, 2010; Pava-Ripoll mopathogens as endophytes from different plant species (Vega et al., 2011). Despite these newly discovered attributes, attention et al., 2008). Some of these fungi have been reported as naturally has mainly focused on developing these entomopathogenic fungal occurring endophytes, while others have been introduced into species as inundative biopesticides against insect and other arthro- the plant using different inoculation techniques (reviewed in pod pests (de Faria and Wraight, 2007). Inundative releases of bio- Table 1 in Vega, 2008). Pioneering work on entomopathogenic control agents normally rely on the direct action of the released endophytes was conducted with Beauveria bassiana (Balsamo) agents, but not on successive generations (Vincentetal.,2007). Vuillemin (Ascomycota: Hypocreales), a ubiquitous soil-borne fun- However, the emerging multiple roles played by fungal ento- gus that infects a wide range of different insects (>700 insect spe- mopathogens provide promising potential for their indirect, multi- cies; Inglis et al., 2001) and is one of the most commercialized faceted and cost-effective use in sustainable agriculture, for fungal biopesticides (de Faria and Wraight, 2007). Lewis and instance as biofertilizers (Kabaluk and Ericsson, 2007; Sasan and Cossentine (1986) credited the season-long suppression of the Bidochka, 2012; Jaber and Enkerli, 2016, 2017), vertically- European corn borer Ostrinia nubilalis (Hübner) (Lepidoptera: transmitted fungal endophytes (Quesada-Moraga et al., 2014; Pyralidae) in maize Zea mays L. (Poaceae), measured as reduced Lefort et al., 2016), and dual microbial control agents of plant dis- tunneling by the insect, to the establishment of B. bassiana as an eases and arthropod pests (Vega et al., 2009; Ownley et al., 2010; endophyte following application of an aqueous suspension of the Lacey et al., 2015). This article reviews the literature currently avail- fungus to the plants. Subsequent work by Lewis and colleagues able on the endophytic colonization of different host plants by fungal using the same model system indicated successful re-isolation of entomopathogens, and summarizes the negative effects of such col- B. bassiana from internal plant tissues after application of the fun- onization on insect pests and plant pathogens that have been gus using different inoculation methods and examined the in reported to date. It also addresses the possible mechanisms of pro- planta growth and movement of the fungus (reviewed in Arnold tection conferred by endophytic fungal entomopathogens, and and Lewis, 2005). explores the potential use of these fungi as dual microbial control In addition to maize, a wide variety of host plants (including agents against both insect and pathogen pests. Moreover, interac- both agronomic and weed speies) have also been shown to harbour tions amongst endophytic fungal entomopathogens and other endo- B. bassiana as an endophyte (summarized in Table 1). In contrast to phytes are discussed. Finally, current limitations and future research B. bassiana, the host plant range of other fungal entomopathogens directions for the innovative use of endophytic fungal ento- is still growing. For instance, Verticillium (=Lecanicillium) lecanii mopathogens as dual microbial control agents are summarized. (Zimm.) Viegas has been reported as a natural endophyte in bear- berry Arctostaphylos uva-ursi L. (Ericaceae) (Widler and Muller, 2. Fungal entomopathogens as endophytes 1984) and ironwood (Bills and Polishook, 1991). Similarly, several genera of known entomopathogenic fungi, including Acremonium, The term endophyte, as introduced by de Bary (1866), broadly Cladosporium, Clonostachys, and Paecilomyces, have been more refers to any organism found within tissues of living autotrophs. recently isolated as naturally-occurring endophytes from various

Table 1 genome-endophyte genome interaction. Leaf surface chemistry Summary of host plants in which Beauveria bassiana (Balsamo) Vuillemin (Ascomy- (Griffin, 2007; Posada et al., 2007) and competition with other cota: Hypocreales) has been reported as an endophyte. endophytes naturally occurring within plants (Posada et al., Host plant References 2007; Schulz et al., 2015; Jaber and Enkerli, 2016) could also lead Bark of ironwood Carpinus caroliniana Walter Bills and Polishook (1991) to differential colonization rates of plants by fungal isolates. (Betulaceae) Indeed, one of the most notable characteristics of fungal ento- Potato Solanum tuberosum L. (Solanaceae); Jones (1994) mopathogens is their diversity in host specificity, ranging from cotton Gossypium hirsutum L. (Malvaceae); very narrow for the obligate pathogens to very wide for the more common cocklebur Xanthium strumarium L. (Asteraceae); jimsonweed Datura facultative ones (Wraight et al., 2007). Several of the genera of fun- stramonium L. (Solanaceae) gal entomopathogens with wide host ranges are also reported to be Cocoa Theobroma gileri Coatrec; Theobroma Evans et al. (2003), Posada endophytes (Vega, 2008; Vega et al., 2008). Such a continuum of cacoa L. (Malvaceae) and Vega (2005) host range, and consequently life history, for the more facultative Seeds and needles of western white pine Pinus Ganley and Newcombe monticola Dougl. ex. D. Don; New Zealand (2005), Reay et al. (2010) entomopathogens could stem from their ability to acquire nutri- pine Pinus radiata D. Don (Pinaceae) ents from sources other than insects (Humber, 2008), which osten- Opium poppy Papaver somniferum L. Quesada-Moraga et al. sibly allows for adaptive inter-kingdom host-jumps from (Papaveraceae) (2006) arthropods to plants and vice versa (Spatafora et al., 2007; Date palm Phoenix dactylifera L. (Arecaceae) Gómez-Vidal et al. (2006) Humber, 2008). Interestingly, Barelli et al. (2016) have recently Banana Musa spp. (Musaceae) Akello et al. (2007) Coffee Coffea arabica L. (Rubiaceae) Posada et al. (2007) suggested that many of these fungi have never left their role as Tomato Lycopersicon esculentum L. Ownley et al. (2008) plant symbionts and hypothesized, as an alternative, that insect (Solanaceae) pathogenicity is an adaptation that allowed certain species of Wheat Triticum aestivum L. (Poaceae); bean Gurulingappa et al. (2010) endophytic fungi to access a specialized source of nitrogen and Phaseolus vulgaris L. (Fabaceae); pumpkin Cucurbita maxima L. (Cucurbitaceae) other nutrients, derived from insects, and effectively exchange Jute Corchorus olitorius L. (Malvaceae) Biswas et al. (2012) these insect-derived nutrients for access to plant carbohydrates. Squash Cucurbita pepo L. (Cucurbitaceae) Jaber and Salem (2014) Alternatively, the bodyguard hypothesis suggests that plants could Artichoke Cynara scolymus L. (Asteraceae) Guesmi-Jouini et al. (2014) alter the behaviour of fungal entomopathogens to increase their Grapevine Vitis vinifera L. (Vitaceae) Jaber (2015) suppression of herbivorous pests and thereby increase plant fitness Oilseed rape Brassica napus L. (Brassicaceae) Vidal and Jaber (2015) Tobacco Nicotiana tabacum L. (Solanaceae); Russo et al. (2015) (Elliot et al., 2000). Fungal entomopathogens spend a significant soybean Glycine max (L.) Merr. (Fabaceae) period of time on the plant surface and are thus vulnerable to plant Cassava Manihot esculenta Crantz Greenfield et al. (2016) chemistry and surface characteristics in addition to exposure to (Euphorbiaceae) damaging UV radiation and adverse changes in microclimate. Shel- Broad bean Vicia faba L. (Fabaceae) Jaber and Enkerli (2016) Sweet pepper Capsicum annum L. (Solanaceae) Jaber and Araj tering these fungi within plants might subsequently add to the (unpublished) novel ways in which plants could manipulate fungal entomopathogens and modify their efficacy (Cory and Ericsson, 2010). Regardless of the specific evolutionary history of endophytic fungal entomopathogens, and given that many fungal ento- tissues of the coffee plant in several countries (see Table 1 in Vega mopathogens in the Hypocreales are generalists with no strict host et al., 2008). preference, it is not a great intuitive leap to suppose that selecting Following foliar spray with conidial suspension, Lecanicillium a virulent isolate capable of extensive endophytic colonization of a lecanii (Zimm.) Zare & Gams and Aspergillus parasiticus Speare have particular host plant would be sufficient to deal with not only its been artificially introduced as endophytes into cotton, wheat, corn, insect pests but also its plant pathogens (Ownley et al., 2010). bean, tomato, and pumpkin (Gurulingappa et al., 2010). Other examples of introduced endophytes include Hypocrea lixii Pat., Gib- berella moniliformis Wineland, Fusarium oxysporum Schlechtendahl 3. Endophytic fungal entomopathogens and potential for emend. Snyder & Hansen and Trichoderma asperellum Samuels, biological control of insect pests Lieckfeldt & Nirenberg into common and broad bean (Akello and Sikora, 2012; Akutse et al., 2013); Purpureocillium lilacinum Thom In addition to their direct biocontrol action against insect pests (formerly Paecilomyces lilacinus) into cotton (Castillo-Lopez et al., through foliar spray or soil application, it has become widely estab- 2014); Clonostachys rosea, Trichoderma harzianum, Trichoderma lished that fungal entomopathogens may also play an important atroviride, T. asperellum, H. lixii and Fusarium sp. into onion Allium role in reducing herbivory following their colonization of plants cepa L. (Amaryllidaceae) (Muvea et al., 2014); Metarhizium robertsii as endophytes. Plant colonization by B. bassiana has been reported (Metchnikoff) Sorokin and Isaria fumosorosea (Wize) Brown & to reduce damage caused by the lepidopteran cob- and stem- Smith into sweet sorghum, Sorghum bicolor (L.) Moench (Poaceae) borers O. nubilalis and Sesamia calamistis Hampson (Lepidoptera: (Mantzoukas et al., 2015); Metarhizium anisopliae (Metchnikoff) Noctuidae) in maize (Bing and Lewis, 1991; Cherry et al., 2004); Sorokin into tomato (Garcia et al., 2011), broad bean (Akello and the tomato fruitworm Helicoverpa zea Boddie (Lepidoptera: Noctu- Sikora, 2012), and cassava (Greenfield et al., 2016); Metarhizium idae) in tomato (Powell et al., 2007); the banana weevil, Cosmopo- pingshaense Q.T. Chen & H.L. Guo into corn (Peña-Peña et al., lites sordidus Germar (Coleoptera: Curculionidae) in banana (Akello 2015); Metarhizium brunneum Petch into broad bean (Jaber and et al., 2008a, 2008b); the poppy stem gall wasp, Iraella luteipes Enkerli, 2016, 2017), potato (Ríos-Moreno et al., 2016), and sweet Thompson (Hymenoptera: Cynipidae) in opium poppy (Quesada- pepper (Jaber and Araj, unpublished); and Beauveria brongniartii Moraga et al., 2009); and the stem weevil Apion corchori Marshall (Saccardo) Petch into broad bean (Jaber and Enkerli, 2017). (Coleoptera: Curculionidae) in white jute (Biswas et al., 2013). Failure to establish an endophytic association has also been Negative effects against insect pests have also been reported as a demonstrated for some isolates of fungal entomopathogens result of plant inoculation with other fungal entomopathogens in (Akutse et al., 2013; Vidal and Jaber, 2015; Mutune et al., 2016). addition to B. bassiana, such as the cotton aphid, Aphis gossypii Glo- This might be due to innate characteristics of the fungal isolate ver (Hemiptera: Aphididae) and the Australian plague locust, Chor- (Posada et al., 2007) or host plant genetics (Arnold and Lewis, toicetes terminifera Walker (Orthoptera: Acrididae) with L. lecanii 2005) leading to potentially unique outcomes for each plant and A. parasiticus in cotton and wheat (Gurulingappa et al.,

2010); the pea aphid, Acyrthosiphon pisum Harris (Hemiptera: tified the fungal secondary metabolites produced in plant tissues Aphididae); the black bean aphid, Aphis fabae Scopoli (Hemiptera: colonized by entomopathogenic fungi. For example, in planta pro- Aphididae) and the pea leafminer, Liriomyza huidobrensis Blanchard duction of destruxins (DTXs) was measured in cowpea plants (Diptera: Agromyzidae) with H. lixii, G. moniliformis, F. oxysporum endophytically-colonized by M. robertsii ARSEF 2575 twelve days and T. asperellum in broad bean (Akello and Sikora, 2012; Akutse after fungal inoculation (Golo et al., 2014). Similarly, destruxin A et al., 2013); onion thrips, Thrips tabaci Lindeman (Thysanoptera: was quantified in melon (Garrido-Jurado et al., 2017) and potato Thripidae) with C. rosea, H. lixii, T. harzianum, T. asperellum, T. atro- (Ríos-Moreno et al., 2016) leaves inoculated by several strains of viride and Fusarium sp. in onion (Muvea et al., 2014); A. gossypii and M. brunneum 72 and 96 h post-inoculation, respectively. The latter the cotton bollworm, Helicoverpa zea Boddie (Lepidoptera: Noctu- study has however reported that the amount of destruxin A pro- idae) with P. lilacinum in cotton (Castillo-Lopez et al., 2014; duced by M. brunneum within plant tissues was very small com- Lopez and Sword, 2015); the Mediterranean corn stalk borer, Sesa- pared to the degree of plant colonization by the fungus, mia nonagrioides Lefebre (Lepidoptera: Noctuidae) with M. robertsii indicating that destruxin A production by the fungus in planta and I. fumosorosea in sweet sorghum (Mantzoukas et al., 2015); the might only be ephemeral (Ríos-Moreno et al., 2016). The exact white grub, Anomala cincta Say (Coleoptera: Melolonthidae) with mode of action by which endophytic fungal entomopathogens con- M. pingshaense in corn (Peña-Peña et al., 2015); the bean stem fer protection against herbivory must be fully elucidated before maggot, Ophiomyia phaseoli Tryon (Diptera: Agromyzidae) with these fungi can be effectively used as biocontrol agents for the M. anisopliae, H. lixii, T. asperellum and T. atroviride in bean management of insect pests. (Mutune et al., 2016); and the green peach aphid, Myzus persicae Sulzer (Hemiptera: Aphididae) with M. brunneum in sweet pepper (Jaber and Araj, unpublished). 4. Endophytic fungal entomopathogens and potential for Recent work shows that transient endophytic colonization by B. biological control of plant diseases bassiana and M. brunneum, following the foliar application of conidia may cause additional mortality in the larvae of the beet army- There is now substantial evidence that some endophytic fungal worm Spodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae) entomopathogens, particularly B. bassiana and Lecanicillium spp. (Resquín-Romero et al., 2016) and nymphs of the sweet potato (formerly Verticillium lecanii), may also demonstrate antagonistic whitefly Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) activity against plant pathogens in addition to their well-known (Garrido-Jurado et al., 2017). Thus, the combined action of foliar biocontrol activity against insect pests. This suggests that these sprays and endophytic colonization could improve the overall effi- entomopathogens have a promising potential to be developed as cacy of commercially–available mycopesticides based on these biopesticides for multiple purposes in IPM strategies (Goettel fungi. It could also, more importantly, help overcome some of the et al., 2008; Vega et al., 2009; Ownley et al., 2010). Beauveria bassi- constraints associated with the topical application of mycopesti- ana strain 11–98, applied as a seed treatment, has been reported to cides, such as inoculum or dosage selection, adverse effects of abi- suppress damping-off caused by the soil-borne pathogens, Rhizoc- otic factors and potential effects on non-target organisms (Lacey tonia solani Kuhn (Basidiomycota: Cantharellales) and Pythium et al., 2015). Alternatively, the underlying mechanisms for the myriotylum Drechsler (Oomycota: Pythiales), in tomato (Ownley detrimental effects against herbivores feeding on plants harbour- et al., 2004; Clark et al., 2006) and cotton seedlings (Griffin, ing fungal entomopathogens as endophytes remain unconfirmed 2007; Ownley et al., 2008). Pre-treatment of cotton seedlings with (Vidal and Jaber, 2015). Mycosis of insect cadavers recovered from the same B. bassiana strain also resulted in reduced severity of bac- plant tissues colonized with fungal entomopathogens has only terial blight caused by Xanthomonas axonopodis pv. malvacearum been reported in a very small number of studies (e.g., Powell (Xam) (Griffin et al., 2006; Ownley et al., 2008). More recently, sev- et al., 2007; Akello et al., 2008b; Vidal and Jaber, 2015; Garrido- eral strains of B. bassiana, including strain ATCC 74040 which is the Jurado et al., 2017). In some cases, the negative effects of endo- active ingredient of the formulated product NaturalisÒ, were found phytic fungal entomopathogens on herbivorous insects have been to significantly reduce the incidence and severity of the Zucchini attributed to induced systemic plant resistance (Akello and yellow mosaic virus (ZYMV; genus Potyvirus, family Potyviridae) Sikora, 2012; Martinuz et al., 2012; Castillo-Lopez et al., 2014; in squash (Jaber and Salem, 2014) and downy mildew caused by Lopez and Sword, 2015; Jaber and Araj, unpublished). However, Plasmopara viticola (Berk. and Curt.) Berl. and de Toni. (Oomycota: specific mechanisms for systemically-induced resistance have not Peronosporaceae) in grapevines (Jaber, 2015) following foliar inoc- been investigated in any of these studies. While systemic plant ulation of plants with conidial suspensions of the tested strains. In resistance has not yet been described against herbivorous insects, contrast, previous studies have mostly investigated the antagonis- it has already been described for resistance to fungal and bacterial tic activity of isolates of Lecanicillium spp. against plant pathogens plant diseases induced by fungal entomopathogens such as Beau- using laboratory, rather than in planta bioassays (see Table 2 in veria bassiana and Lecanicillium species (discussed in Section 5 Ownley et al., 2010 for a review of these studies). To date, only a below); it is possible that similar systemic resistance is elicited handful of studies have reported biological control of plant patho- by these fungi against insect herbivores as well. Notably, feeding gens by endophytic Lecanicillium spp. For example, induced resis- deterrence or antibiosis due to fungal metabolites secreted in tance against Pythium ultimum Trow (Oomycota: Pythiales) planta has been widely suggested as the mode of action in several (Benhamou and Brodeur, 2001) and powdery mildew Sphaerotheca studies investigating endophytic entomopathogenic fungi- fuliginea (Schlecht.: Fr.) Pollacci (Leotiomycetes: Erysiphaceae) herbivorous insect interactions (e.g., Lewis and Cossentine, 1986; (Hirano et al., 2008) have been attributed to limited endophytic Bing and Lewis, 1991; Cherry et al., 2004; Akello et al., 2008a; colonization of cucumber roots by Lecanicillium sp. DAOM Quesada-Moraga et al., 2009; Gurulingappa et al., 2010; Akutse 198499 and Lecanicillium muscarium B-2, respectively. In a study et al., 2013; Castillo-Lopez et al., 2014; Golo et al., 2014; Muvea that coupled both in vitro and in vivo bioassays, endophytic et al., 2014; Mantzoukas et al., 2015; Garrido-Jurado et al., 2017; M. robertsii conferred protection against root rot of bean caused Lefort et al., 2016; Mutune et al., 2016; Ríos-Moreno et al., 2016). by Fusarium solani (Mart.) Sacc. f. sp. phaseoli (Burkholder) W.C. This suggestion has been supported by the absence of fungal Snyder & N.H. Hans (Ascomycota: Nectriaceae) (Sasan and sporulation (mycosis) on insects that have died when feeding on Bidochka, 2013). Coupling in vitro, in vivo, and field tests in differ- endophytically-colonized plants by fungal entomopathogens. ent environments is the most robust approach to screen fungal Despite this, only a few of these studies have identified and quan- endophytes for disease suppression (Busby et al., 2016). Most

Please cite this article in press as: Jaber, L.R., Ownley, B.H. Can we use entomopathogenic fungi as endophytes for dual biological control of insect pests and plant pathogens?. Biological Control (2017), http://dx.doi.org/10.1016/j.biocontrol.2017.01.018 L.R. Jaber, B.H. Ownley / Biological Control xxx (2017) xxx–xxx 5 recently, an endophytic isolate of Phialemonium inflatum (formerly has successfully established within plant tissues, the fungus occu- Paecilomyces inflatus) has been reported to suppress penetration, pies a niche and depends on nutrients provided by its host plant. galling and reproduction of the root-knot nematode Meloidogyne Given that the fungal endophyte is the first to colonize the plant, incognita (Kofoid & White) Chitwood (Tylenchida: Meloidogy- it is likely that it exhausts the host plant resources leaving none nidae) in cotton plants following seed treatment with the conidial available for the plant pathogen when it attempts to colonize. suspension of the fungus overnight (Zhou et al., 2016). This study Change in the order of plant colonization, for example when the presents the first evidence for the potential use of ento- pathogen colonizes the plant before the endophyte, can shift the mopathogenic and nematophagous fungi for the biological control endophyte-pathogen interaction from disease suppression to dis- of phytoparasitic nematodes following their endophytic coloniza- ease facilitation (see Adame-Álvarez et al., 2014). Initial endo- tion of plants. phytic colonization also induces plants to produce lignin and other cell wall deposits as a mechanical defense response and this too might consequently prevent or limit infection by disease- 5. Possible mechanisms of plant disease control by endophytic causing plant pathogens (see Schulz and Boyle, 2005). fungal entomopathogens Antibiosis, by the production of secondary metabolites, has been reported as one of the mechanisms by which fungal ento- Although a growing number of studies have recently demon- mopathogens confer protection against disease-causing plant strated that ‘fungal entomopathogens as endophytes’ can protect pathogens and insect pests (Ownley et al., 2010). Fungal ento- their host plant against plant pathogens or limit damaging effects, mopathogens are a rich source of secondary metabolites with our knowledge about the precise mechanism(s) underlying such antimicrobial, insecticidal, and cytotoxic activities (Gibson et al., endophytic protection by these fungi remains at an early stage. 2014). For instance, B. bassiana produces numerous secondary Here we discuss potential mechanisms, ranging from those directly metabolites including beauvericin, bassianin, beauverolides, affecting plant pathogens (e.g., mycoparasitism, competition, bassianolides, oosporein, bassianolone amongst others (reviewed antibiosis through production of fungal secondary metabolites) to in Ownley et al., 2010). Of these compounds, beauvericin in partic- the more complex indirect interactions between endophytic fungal ular has broad and significant multiple bioactivities and can also be entomopathogens and disease causing pathogens as mediated produced by several entomopathogenic fungal genera, such as Pae- through their mutual host plant (e.g., induction of systemic plant cilomyces, Isaria and Fusarium (Wang and Xu, 2012). Beauvericin is resistance, stimulation of plant secondary metabolites and promo- produced during broth culture by B. bassiana strain 11–98 (Leckie tion of plant growth). Strong evidence suggests that a combination et al., 2008), an endophytic strain found to suppress damping-off of these mechanisms, rather than a single mechanism, might be caused by R. solani and P. myriotylum in tomato and cotton employed by endophytic fungal entomopathogens against plant (Ownley et al., 2004, 2008; Clark et al., 2006; Griffin, 2007). How- pathogens (Vega et al., 2009; Ownley et al., 2010). ever, this fungal secondary compound remains to be detected within plants colonized by B. bassiana at the concentrations neces- 5.1. Direct suppression of plant pathogens sary to suppress the respective fungal plant pathogens in planta. Similarly, the production of several other secondary metabolites Endophytic fungal entomopathogens can directly suppress with antimicrobial activities by endophytic entomopathogenic plant pathogens through mycoparasitism, competition for ecologi- fungi has hitherto only been reported in in vitro studies (e.g., cal niches and nutrition, and by production of secondary metabo- Wainwright et al., 1986; Lee et al., 2005; Carollo et al., 2010; lites. Mycoparasitism, defined as the interrelationship between a Sasan and Bidochka, 2013). fungus parasite and a fungus host (Barnett, 1963; Jeffries, 1995), has been extensively studied for some entomopathogenic fungi, 5.2. Induction of systemic plant resistance particularly species in the genera Trichoderma (Steyaert et al., 2003; Harmon et al., 2004) and Lecanicillium (see Ownley et al., Induced systemic resistance (ISR), known to be elicited by ben- 2010; and references therein). It is mainly characterized by the for- eficial microbes, has emerged as an important mechanism by mation of coiled hyphal structures around the hyphae of the host which the whole plant is primed for enhanced defense against a fungus, which has been observed for an endophytic strain of B. broad range of plant pathogens and insects pests (Pieterse et al., bassiana in parasitism assays against P. myriotylum (Griffin, 2014). Evidence for ISR includes reduction of disease symptoms 2007). Mycoparasitism also involves penetration of the cell wall in plant parts distant from the site where the inducing agent is due to the production of lytic enzymes that break down cell wall active. This has been shown in cotton seedlings inoculated with components (Askary et al., 1997; Zeilinger et al., 1999), and the B. bassiana strain 11–98 using a root drench followed by foliar chal- release of antibiotics that permeate the perforated hyphae and pre- lenge with the bacterial blight pathogen Xanthomonas 13 days vent re-synthesis of the host cell wall (Askary et al., 1997; Lorito later. Root inoculation with B. bassiana resulted in significantly et al., 1996) and growth of the host hyphal cytoplasm (Inbar reduced disease severity ratings for bacterial blight on the leaves et al., 1996). To date, the process of mycoparasitism by fungal ento- of inoculated plants compared with the untreated control plants. mopathogens (e.g., Lecanicillium spp. and Trichoderma spp.) has Furthermore, B. bassiana inoculation was as effective as treatment been thoroughly described in vitro under laboratory conditions, with 2,6-dichloro-isonicotinic acid, a chemical known to induce but remains to be demonstrated in planta. systemic resistance against plant pathogens (Griffin et al., 2006; Competition for space and nutrition is likely to be the biocontrol Griffin, 2007; Ownley et al., 2008). Similarly, ISR has been reported mechanism operating against R. solani (Ownley et al., 2004; Griffin, against P. ultimum (Benhamou and Brodeur, 2001) and powdery 2007) and P. viticola (Jaber, 2015)inB. bassiana endophytically- mildew S. fuliginea (Hirano et al., 2008) as a result of pre- colonized plants. Plant colonization by B. bassiana was confirmed inoculation of cucumber roots with Lecanicillium spp. Colonization in grapevine plants with reduced incidence and severity of downy of date palm by B. bassiana and Lecanicillium spp. resulted in the mildew symptoms before the disease-causing pathogen (P. viticola) up-regulation of proteins involved in plant defense and stress had been introduced into plants (Jaber, 2015). Colonization of plant response (Gómez-Vidal et al., 2009), thus inducing a ‘primed state’ tissues by fungal endophytes involves several steps including host by which plants would be able to achieve both faster and stronger recognition, spore germination, penetration of the plant surface activation of a broad-spectrum resistance to pathogens, insects, and tissue colonization (Petrini, 1991). Once fungal colonization and abiotic stress (Conrath et al., 2006). Suppression or delay of

Endophytic fungal entomopathogens may contribute to protec- Consortia of endophytic microorganisms have been studied for tion of their host plant against disease pathogens through simultaneous management of pests and pathogens, in an effort to enhancement of plant growth. A growing number of studies have increase the variety of effective modes of action and expand the lately demonstrated the ability of several fungal entomopathogens range of pests and pathogens that are negatively impacted by these to promote plant growth following endophytic establishment (e.g., bioformulations. Compatible consortia have been reported for syn- Garcia et al., 2011; Sasan and Bidochka, 2012; Liao et al., 2014; chronous control of sheath blight disease (R. solani) and leaffolder Lopez and Sword, 2015; Jaber and Enkerli, 2016, 2017; Jaber and insect (Cnaphalocrocis medinalis Guen.) on rice (Karthiba et al., Araj, unpublished). Increased plant growth, mediated by coloniza- 2009). The consortium included two strains of plant growth- tion by fungal endophytes, results in suppression of various abiotic promoting rhizobacteria (Pseudomonas fluorescens Migula) and B. and biotic stresses, including plant diseases (Kuldau and Bacon, bassiana strain B2. An aqueous talc-based formulation of all three 2008). This has been shown in B. bassiana-colonized squash plants organisms, which was applied three times; to seed, as a root dip against ZYMV (Jaber and Salem, 2014). When challenged with to seedlings 25 days after planting and as a foliar spray 30 days ZYMV, B. bassiana-inoculated plants not only expressed reduced after planting, significantly reduced the incidences of both pest disease incidence and severity, but also appeared more vigorous and disease, and increased growth promotion of rice and of grain and developed faster than the non-inoculated control plants. The yield. Tissues of the treated plants also accumulated greater same has been demonstrated in plants colonized by M. robertsii amounts of defense enzymes, such as chitinase, lipoxygenase, per- and exposed to F. solani; the M. robertsii–inoculated plants showed oxidase, and polyphenol oxidase, compared with untreated healthier growth and lower disease indices compared with plants controls. not colonized by M. robertsii (Sasan and Bidochka, 2013). Indeed, A similar combination (strains of B. bassiana and P. fluorescens) plant inoculation with fungal entomopathogens (e.g., B. bassiana significantly reduced damage caused by leafminer (Aproaerema and Lecanicillium spp.) have induced proteins related to photosyn- modicella Deventer) and collar rot (Sclerotium rolfsii Sacc.) on thesis and energy metabolism as well as plant defense and groundnut (Arachis hypogaea L.) (Senthilraja et al., 2010a). This responses to stress, which could enhance plant growth and stimu- microbial combination was applied as a talc-based formulation to late disease resistance (Gómez-Vidal et al., 2009). Enhanced plant seed, soil, and foliage. Efficacy against pest and disease, as well growth observed in plants colonized by fungal entomopathogens as yield of groundnut, was further increased when chitin was might also be attributed to the production of phytohormones or added to the bioformulation (Senthilraja et al., 2010b). In both siderophores (i.e., low molecular weight iron-chelating compounds glasshouse and field, the consortium performed significantly better synthesized by microorganisms). For example, M. anisopliae has than individual components of the bioformulation, with or without been shown to elicit production of phytohormones in inoculated chitin, or standard pesticide treatments (Senthilraja et al., 2010b). soybean plants (Khan et al., 2012), whereas M. robertsii and B. A consortium of two endophytic fungi, B. bassiana strain 11–98 bassiana have been found to produce siderophores under and an arbuscular mycorrhizal fungus, R. intraradices, was

Please cite this article in press as: Jaber, L.R., Ownley, B.H. Can we use entomopathogenic fungi as endophytes for dual biological control of insect pests and plant pathogens?. Biological Control (2017), http://dx.doi.org/10.1016/j.biocontrol.2017.01.018 L.R. Jaber, B.H. Ownley / Biological Control xxx (2017) xxx–xxx 7 evaluated for effects on secondary metabolites in tomato plants as a complementary tool in IPM programmes. However, fungal and on defense against beet armyworm (Spodoptera exigua Hilb- entomopathogens may not fully colonize all plant tissues or persist ner) (Shrivastava et al., 2015). Control plants, which were not inoc- for long periods of time due to multipartite interactions with other ulated with either fungus, constitutively produced monoterpenes bacterial and fungal inhabitants within the host plants (see Schulz and sesquiterpenes. However, inoculation with the two endo- et al., 2015). A deeper knowledge about the extent and persistence phytes, singly or combined, increased the amount of monoterpenes of entomopathogenic fungi inside plants is required and consti- and sesquiterpenes detected, and revealed new monoterpenes not tutes the basis for determining the degree of plant protection as found in control plants. In feeding assays with beet armyworm, lar- well as other benefits conferred by these fungi as endophytes. Pre- vae fed on tomato plants inoculated with either or both fungal spe- vious studies have reported the endophytic colonization of plants cies gained significantly less weight than those fed on control non- by B. bassiana to continue for as long as three months in jute inoculated plants. This suggests that the defense response of (Biswas et al., 2012), eight months in coffee (Posada et al., 2007) fungus-inoculated plants was stronger than control plants, which and nine months in radiata pine (Brownbridge et al., 2012). Colo- may be partly due to the difference in levels of terpenoids. nization levels of cassava by M. anisopliae remained constant from 7–9 dpi (80%) to 47–49 dpi (80%), whereas colonization levels of B. bassiana decreased from 83% at 7–9 dpi to 40% at 47–49 dpi 7. Conclusions and future prospects (Greenfield et al., 2016). Under field conditions, B. bassiana, M. robertsii, and I. fumosorosea were re-isolated 30 days after foliar Due to increased environmental concerns and pest resistance spray of sweet sorghum (Mantzoukas et al., 2015). The extent problems, biopesticides have been viewed as a promising replace- and persistence of endophytic fungal colonization within plants ment for synthetic pesticides and a key component of environmen- can be improved by repeated application of the microbial agent tally friendly pest management (Glare et al., 2012). through foliar spray or soil drench (Jaber unpublished). Impor- Entomopathogenic fungi are a unique and highly specialized group tantly, researchers should be aware of the fact that serial sub- of microbial agents that possess several desirable traits favouring culturing on artificial media reduces viability and virulence, which their development as biopesticides (Lacey et al., 2015). Although may therefore alter functionality of fungal isolates including their there are almost 700 species in about 100 genera of fungal ento- endophytic capacity to colonize plants. Storing agar plugs of fungal mopathogens (Humber, 2008), the majority of the commercially isolates in tubes of sterile distilled water can maintain viability produced fungi are only based on a few species of Beauveria, over long periods of time (Richter et al., 2016). Alternatively, pas- Metarhizium, Isaria, and Lecanicillium (see Table 1 in de Faria and sage through insect hosts (Nahar et al., 2008) or re-isolation of Wraight, 2007 and Table 3 in Lacey et al., 2015). Indeed, a better endophytes from inoculated plants (Busby et al., 2016) may serve understanding of the ecology of fungal entomopathogens would to refresh the ecological function of fungal isolates as endophytes. stimulate the development and uptake of more commercially- Fungal entomopathogens may colonize particular host plants available biopesticides based on these fungi in mainstream agricul- more efficiently than others, which might consequently influence ture (Vega et al., 2009; Glare et al., 2012; Lacey et al., 2015). Fur- the level of plant protection by the colonizing fungi. The inocula- thermore, to encourage the widespread use of fungal tion method is shown to affect the efficiency of colonization and entomopathogen-based biopesticides there is a need for products high inoculum rates are usually optimal for enhancing colonization with activity against multiple pests in addition to improved deliv- levels, suppressing insect and pathogen pests and providing plant ery methods and increased persistence (Glare et al., 2012). More growth promotion benefits (e.g., Posada et al., 2007; Akello et al., evaluation under field conditions is required and increased atten- 2009; Tefera and Vidal, 2009; Garcia et al., 2011; Muvea et al., tion must be paid to ensure compatibility and maximize efficacy 2014; Russo et al., 2015; Jaber and Enkerli, 2016, 2017). The extent when trying to incorporate fungal entomopathogen-based biopes- of endophytic plant colonization may also vary depending on the ticides within IPM programmes (Lacey et al., 2015). methodology used for detecting the endophytic organism within Endophytes are pesticidal microbes for which the plant acts as a plants (Hyde and Soytong, 2008; McKinnon et al., 2017). Future delivery system; this fits the definition of a biopesticide (see Box 1 studies are therefore recommended to employ a combination of in Glare et al., 2012). Fungal entomopathogens as endophytes are culture-dependent (i.e., direct isolation of endophytic fungal mate- rapidly becoming a distinct group of microbial biocontrol agents rial from within plant tissues onto a growth media) and culture- as indicated by the recent surge of published studies exploring independent (i.e., molecular detection of endophytic fungal DNA the endophytic potential of entomopathogenic fungi. The use of within plant tissues by PCR) techniques for more reliable detection endophytic fungal entomopathogens as seed treatments intro- of endophytic plant colonization by fungal entomopathogens duced at an early stage of plant development overcomes several (Biswas et al., 2012; Jaber, 2015; McKinnon et al., 2017). Research- inherent problems usually encountered when using fungal ento- ers should also realize the importance of the surface-sterilization mopathogens as contact biocontrol agents. These include exposure methods they use for recovering endophytes and eliminating epi- to detrimental environmental conditions (e.g., damaging UV radia- phytes. Making imprints of surface-sterilized tissue on the agar tion, reduced humidity, and excessive rainfall), compatibility with media, as introduced by Schultz et al. (1998), has proved to be other control measures, and the challenge of synchronizing the more reliable than plating of final rinse water for testing the effi- biocontrol agent with target pests. In addition to their promising cacy of the surface-sterilization protocol used for isolating endo- dual biocontrol potential against insect pests and plant diseases, phytes (Hyde and Soytong, 2008; McKinnon et al., 2017). fungal entomopathogens as endophytes may also offer protection Research efforts should now focus on controlling the potential against cryptic pests (e.g., insect borers) that would otherwise be effects of fungal genotype-plant genotype interactions. Screening difficult to control by topical application (Jaronski, 2010). They strains or isolates in order to select the ones most adapted to endo- can also provide additional benefits such as accelerating seedling phytism in a specific host plant, or even cultivar, may enhance the emergence and improved plant growth (Sasan and Bidochka, colonization rate of plant tissues. Selecting superior endophytic 2012; Lopez and Sword, 2015; Jaber and Enkerli, 2016, 2017; Jaber strains with high virulence against one or more pests could subse- and Araj, unpublished). Recently, endophytic fungal ento- quently facilitate the development of these strains for wider man- mopathogens have been found to be compatible with other biocon- agement of multiple plant pests. Furthermore, the differential trol agents such as parasitoids (e.g., Akutse et al., 2014; Gathage expression of fungal genotype-plant genotype interactions under et al., 2016; Jaber and Araj, unpublished) and may thus be used different environmental conditions should be taken into account.

Notably, only very few studies have so far investigated endophytic Askary, H., Benhamou, N., Brodeur, J., 1997. Ultrastructural and cytochemical fungal entomopathogens under field conditions (e.g., Cherry et al., investigations of the antagonistic effect of Verticillium lecanii on cucumber powdery mildew. Phytopathology 87, 359–368. 2004; Quesada-Moraga et al., 2009; Castillo-Lopez et al., 2014; Barelli, L., Moonjely, S., Behie, S.W., Bidochka, M.J., 2016. Fungi with multifunctional Mantzoukas et al., 2015; Gathage et al., 2016). In order to translate lifestyles: endophytic insect pathogenic fungi. Plant Mol. Biol. 90, 657–664. the promising biocontrol efficacy of many endophytic ento- Barnett, H.L., 1963. The nature of mycoparasitism by fungi. Annu. Rev. Microbiol. 17, 1–44. mopathogenic fungal isolates tested under controlled experimen- Behie, S.W., Bidochka, M.J., 2014. An additional branch of the soil nitrogen cycle: tal conditions into a similar efficacy under more natural field ubiquity of insect-derived nitrogen transfer to plants by endophytic insect conditions, special attention should be paid by researchers to pathogenic fungi. Appl. Environ. Microbiol. 80, 1553–1560. Benhamou, N., Brodeur, J., 2001. Pre-inoculation of Ri T-DNA transformed cucumber improve the limited ‘external validity’ that is frequently reported roots with the mycoparasite, Verticillium lecanii, induces host defense reactions by experimental studies trying to extend conclusions to situations against Pythium ultimum infection. Physiol. Mol. Plant Pathol. 58, 133–146. of different environmental conditions (Rosenheim et al., 2011). For Bills, G.F., Polishook, J.D., 1991. Microfungi from Carpinus caroliniana. Can. J. Bot. 69, 1477–1482. example, even though experimental studies that investigate endo- Bing, L.A., Lewis, L.C., 1991. Suppression of Ostrinia nubilalis (Hübner) (Lepidoptera: phytic fungal entomopathogens using sterile planting substrates Pyralidae) by endophytic Beauveria bassiana (Balsamo) Vuillemin. Environ. have merit and thus a strong ‘internal validity’, they will probably Entomol. 20, 1207–1211. have a limited ‘external validity’ because sterile soils do not exist in Biswas, C., Dey, P., Satpathy, S., Satya, P., 2012. Establishment of the fungal entomopathogen Beauveria bassiana as a season long endophyte in jute nature (Parsa et al., 2016). Our challenge as scientists is to pave the (Corchorus olitorius) and its rapid detection using SCAR marker. Biocontrol 57, way for the full potential of fungal entomopathogens as endo- 565–571. phytes to be realized for the integrated management of multiple Biswas, C., Dey, P., Satpathy, S., Satya, P., Mahapatra, B.S., 2013. Endophytic colonization of white jute (Corchorus capsularis) plants by different Beauveria pests. However, several problems mainly regarding the consistency bassiana strains for managing stem weevil (Apion corchori). Phytoparasitica 41, of plant colonization and subsequent endophytic protection need 17–21. to be addressed. There are also many open questions concerning Bourgaud, F., Gravot, A., Milesi, S., Gontier, E., 2001. Production of plant secondary metabolites: a historical perspective. Plant Sci. 161, 839–851. possible modes of action that must be answered before their full Brownbridge, M., Reay, S.D., Nelson, T.L., Glare, T.R., 2012. Persistence of Beauveria potential as effective pest and disease management tools is bassiana (Ascomycota: Hypocreales) as an endophyte following inoculation of achieved. radiata pine seed and seedlings. Biol. Control 61, 194–200. Bruck, D.J., 2010. Fungal entomopathogens in the rhizosphere. Biocontrol 55, 103– 112. Acknowledgments Busby, P.E., Ridout, M., Newcombe, G., 2016. Fungal endophytes: modifiers of plant disease. Plant Mol. Biol. 90, 645–655. Carollo, C.A., Calil, A.L.A., Schiave, L.A., Guaratini, T., Roberts, D.W., Lopes, N.P., Braga, We would like to thank Dr. Judith Pell and Dr. Fernando Vega G.U., 2010. Fungal tyrosine betaine, a novel secondary metabolite from conidia for inviting us to contribute to this Special Issue of Biological Con- of entomopathogenic Metarhizium spp. fungi. Fungal Biol. 114, 473–480. Castillo-Lopez, D., Zhu-Salzman, K., Ek-Ramos, M.J., Sword, G.A., 2014. The trol. This work was supported by the Department of Plant Protec- entomopathogenic fungal endophytes Purpureocillium lilacinum (formerly tion at the University of Jordan and the Department of Paecilomyces lilacinus) and Beauveria bassiana negatively affect cotton aphid Entomology and Plant Pathology at The University of Tennessee. reproduction under both greenhouse and field conditions. PLoS One 9, e103891. Cherry, A.J., Banito, A., Djegui, D., Lomer, C., 2004. Suppression of the stem-borer Sesamia calamistis (Lepidoptera; Noctuidae) in maize following seed dressing, References topical application and stem injection with African isolates of Beauveria bassiana. Int. J. Pest Manage. 50, 67–73. Clark, M.M., Gwinn, K.D., Ownley, B.H., 2006. Biological control of Pythium Adame-Álvarez, R.M., Mendiola-Soto, J., Heil, M., 2014. Order of arrival shifts myriotylum. Phytopathology 96, S25. endophyte-pathogen interactions in bean from resistance induction to disease Conrath, U., Beckers, G.J.M., Flors, B., Garcıa-Agustın, P., Jakab, G., Mauch, F., facilitation. FEMS Microbiol. Lett. 355, 100–107. Newman, M.-A., Pieterse, C.M.J., Poinssot, B., Pozo, M.J., Pugin, A., Schaffrath Ton, Ahmed, A.S., Sa’nchez, C.P., Candela, M.E., 2000. Evaluation of induction of systemic J., Wendehenne, D., Zimmerli, L., Mauch-Mani, B., 2006. Priming: getting ready resistance in pepper plants (Capsicum annuum)toPhytophthora capsici using for battle. Mol. Plant Microbe Interact. 19, 1062–1071. Trichoderma harzianum and its relation with capsidiol accumulation. Eur. J. Plant Cory, J.S., Ericsson, J.D., 2010. Fungal entomopathogens in a tritrophic context. Pathol. 106, 817–824. Biocontrol 55, 129–145. Akello, J., Dubois, T., Coyne, D., Kyamanywa, S., 2008a. Effect of endophytic de Bary, A., 1866. Morphologie und Physiologie der Pilze, Flechten du Beauveria bassiana on populations of the banana weevil, Cosmopolites sordidus, Myxomyceten. Engelmann, Leipzig. and their damage in tissue-cultured banana plants. Entomol. Exp. Appl. 129, de Faria, M.R., Wraight, S.P., 2007. Mycoinsecticides and mycoacaricides: a 157–165. comprehensive list with worldwide coverage and international classification Akello, J., Dubois, T., Coyne, D., Kyamanywa, S., 2008b. Endophytic Beauveria of formulation types. Biol. Control 43, 237–256. bassiana in banana (Musa spp.) reduces banana weevil (Cosmopolites sordidus) Elliot, S.L., Sabelis, M.W., Janssen, A., van der Geest, L.P.S., Beerling, E.A.M., Fransen, fitness and damage. Crop Prot. 27, 1437–1441. J., 2000. Can plants use entomopathogens as bodyguards? Ecol. Lett. 3, 228–235. Akello, J., Dubois, T., Coyne, D., Kyamanywa, S., 2009. The effects of Beauveria Evans, H.C., Holmes, K.A., Thomas, S.E., 2003. Endophytes and mycoparasites bassiana dose and exposure duration on colonization and growth of tissue associated with an indigenous forest tree, Theobroma gileri, in Ecuador and a cultured banana (Musa sp.) plants. Biol. Control 49, 6–10. preliminary assessment of their potential as biocontrol agents of cocoa diseases. Akello, J., Dubois, T., Gold, C.S., Coyne, D., Nakavuma, J., Paparu, P., 2007. Beauveria Mycol. Progr. 2, 149–160. bassiana (Balsamo) Vuillemin as an endophyte in tissue culture banana (Musa Fraser, R.S.S., 1979. Systemic consequences of the local lesion reaction to tobacco spp.). J. Invertebr. Pathol. 96, 34–42. mosaic virus in a tobacco variety lacking the N gene for hypersensitivity. Akello, J., Sikora, R., 2012. Systemic acropedal influence of endophyte seed Physiol. Plant Pathol. 14, 383–394. treatment on Acyrthosiphon pisum and Aphis fabae offspring development and Ganley, R.J., Newcombe, G., 2005. Fungal endophytes in seeds and needles of Pinus reproductive fitness. Biol. Control 61, 215–221. monticola. Mycol. Res. 110, 318–327. Akutse, K.S., Fiaboe, K.K.M., Van den Berg, J., Ekesi, S., Maniania, N.K., 2014. Effects of Garcia, J.E., Posadas, J.B., Perticari, A., Lecuona, R.E., 2011. Metarhizium anisopliae endophyte colonization of Vicia faba (Fabaceae) plants on the life-history of (Metschnikoff) Sorokin promotes growth and has endophytic activity in tomato leafminer parasitoids Phaedrotoma scabriventris (Hymenoptera: Braconidae) plants. Adv. Biol. Res. 5, 22–27. and Diglyphus isaea (Hymenoptera: Eulophidae). PLoS One 9 (10), e109965. Garrido-Jurado, I., Resquín-Romero, G., Amarilla, S.P., Ríos-Moreno, A., Carrasco, L., Akutse, K.S., Maniania, N.K., Fiaboe, K.K.M., Van Den Berg, J., Ekesi, S., 2013. Quesada-Moraga, E., 2017. Transient endophytic colonization of melon plants Endophytic colonization of Vicia faba and Phaseolus vulgaris (Fabaceae) by by entomopathogenic fungi after foliar application for the control of Bemisia fungal pathogens and their effects on the life-history parameters of Liriomyza tabaci Gennadius (Hemiptera: Aleyrodidae). J. Pest. Sci. 90, 319–330. huidobrensis (Diptera: Agromyzidae). Fungal Ecol. 6, 293–301. Gathage, J.W., Lagat, Z.O., Fiaboe, K.K.M., Akutse, K.S., Ekesi, S., Maniania, N.K., 2016. Arnold, A.E., Lewis, L.C., 2005. Ecology and evolution of fungal endophytes and their Prospects of fungal endophytes in the control of Liriomyza leafminer flies in roles against insects. In: Vega, F.E., Blackwell, M. (Eds.), Insect-Fungal common bean Phaseolus vulgaris under field conditions. Biocontrol 61, 741–753. Associations: Ecology and Evolution. Oxford University Press, New York, pp. Gibson, D.M., Donzelli, B.G., Krasnoff, S.B., Keyhani, N.O., 2014. Discovering the 74–96. secondary metabolite potential encoded within entomopathogenic fungi. Nat. Arnold, A.E., Lutzoni, F., 2007. Diversity and host range of foliar fungal endophytes: Prod. Rep. 31, 1287–1305. are tropical leaves biodiversity hotspots? Ecology 88, 541–549. Glare, T.R., Caradus, J., Gelernter, W., Jackson, T., Keyhani, N., Kohl, J., Marrone, P., Arnold, A.E., Maynard, Z., Gilbert, G.S., 2001. Fungal endophytes in dicotyledonous Morin, L., Stewart, A., 2012. Have biopesticides come of age? Trends Biotechnol. neotropical trees: patterns of abundance and diversity. Mycol. Res. 105, 1502– 30, 250–258. 1507.

Goettel, M.S., Koike, M., Kim, J.J., Aiuchi, D., Shinya, R., Brodeur, J., 2008. Potential of Kuldau, G., Bacon, C., 2008. Clavicipitaceous endophytes: their ability to enhance Lecanicillium spp. for management of insects, nematodes and plant diseases. J. resistance of grasses to multiple stresses. Biol. Control 46, 57–71. Invertebr. Pathol. 98, 256–261. Lacey, L.A., Grzywacz, D., Shapiro-Ilan, D.I., Frutos, R., Brownbridge, M., Goettel, M.S., Golo, P.S., Gardner, D.R., Grilley, M.M., Takemoto, J.Y., Krasnoff, S.B., et al., 2014. 2015. Insect pathogens as biological control agents: back to the future. J. Production of destruxins from Metarhizium spp. fungi in artificial medium and Invertebr. Pathol. 132, 1–41. in endophytically colonized cowpea plants. PLoS One 9 (8), e104946. Leckie, B.M., Ownley, B.H., Pereira, R.M., Klingeman, W.E., Jones, C.J., Gwinn, K.D., Gómez-Vidal, S., Lopez-Llorca, L.V., Jansson, H.-B., Salinas, J., 2006. Endophytic 2008. Mycelia and spent fermentation broth of Beauveria bassiana incorporated colonization of date palm (Phoenix dactylifera L.) leaves by entomopathogenic into synthetic diets affect mortality, growth and development of larval fungi. Micron 37, 624–632. Helicoverpa zea (Lepidoptera: Noctuidae). Biocontrol Sci. Technol. 18, 697–710. Gómez-Vidal, S., Salinas, J., Tena, M., Lopez-Llorca, L.V., 2009. Proteomic analysis of Lee, S.-Y., Nakajima, I., Ihara, F., Kinoshita, H., Nihira, T., 2005. Cultivation of date palm (Phoenix dactylifera L.) responses to endophytic colonization by entomopathogenic fungi for the search of antibacterial compounds. entomopathogenic fungi. Electrophoresis 30, 2996–3005. Mycopathologia 160, 321–325. Greenfield, M., Gómez-Jiménez, M.I., Ortiz, V., Vega, F.E., Kramer, M., Parsa, S., 2016. Lefort, M.-C., McKinnon, A.C., Nelson, T.L., Glare, T.R., 2016. Natural occurrence of Beauveria bassiana and Metarhizium anisopliae endophytically colonize cassava the entomopathogenic fungi Beauveria bassiana as a vertically transmitted roots following soil drench inoculation. Biol. Control 95, 40–48. endophyte of Pinus radiate and its effect on above- and below-ground insect Griffin, M.R., 2007. Beauveria Bassiana, A Cotton Endophyte With Biocontrol pests. NZ Plant Prot. 69, 68–77. Activity Against Seedling Disease (Ph.D. Dissertation). The University of Loebenstein, G., 1972. Localization and induced resistance in virus-infected plants. Tennessee, Knoxville, TN, USA. Annu. Rev. Phytopathol. 10, 177–206. Griffin, M.R., Ownley, B.H., Klingeman, W.E., Pereira, R.M., 2006. Evidence of induced Lewis, L.C., Cossentine, J.E., 1986. Season long intraplant epizootics of systemic resistance with Beauveria bassiana against Xanthomonas in cotton. entomopathogens, Beauveria bassiana and Nosema pyrausta, in a corn Phytopathology 96, S42. agroecosystem. Entomophaga 31, 36–69. Guesmi-Jouini, J., Garrido-Jurado, I., López-Díaz, C., Ben Halima-Kamel, M., Quesada- Liao, X., O’Brien, T.R., Fang, W., St. Leger, R.J., 2014. The plant beneficial effects of Moraga, E., 2014. Establishment of fungal entomopathogens Beauveria bassiana Metarhizium species correlate with their association with roots. Appl. Microbiol. and Bionectria ochroleuca (Ascomycota: Hypocreales) as endophytes on Biotechnol. 98, 7089–7096. artichoke Cynara scolymus. J. Invertebr. Pathol. 119, 1–4. Lopez, D.C., Sword, G.A., 2015. The endophytic fungal entomopathogens Beauveria Gurulingappa, P., Sword, G.A., Murdoch, G., McGee, P.A., 2010. Colonization of crop bassiana and Purpureocillium lilacinum enhance the growth of cultivated cotton plants by fungal entomopathogens and their effects on two insect pests when in (Gossypium hirsutum) and negatively affect survival of the cotton bollworm planta. Biol. Control 55, 34–41. (Helicoverpa zea). Biol. Control 89, 53–60. Harmon, G.E., Howell, C.R., Viterbo, A., Chet, I., Lorito, M., 2004. Trichoderma Lorito, M., Farkas, V., Rebuffat, S., Bodo, B., Kubicek, C.P., 1996. Cell wall synthesis is species–opportunistic, avirulent plant symbionts. Nat. Rev. Microbiol. 2, a major target of mycoparasitic antagonism by Trichoderma harzianum.J. 43–56. Bacteriol. 178, 6382–6385. Hartley, S.E., Eschen, R., Horwood, J.M., Gange, A.C., Hill, E.M., 2015. Infection by a Mantzoukas, S., Chondrogiannis, C., Grammatikopoulos, G., 2015. Effects of three foliar endophyte elicits novel arabidopside-based plant defence reactions in its endophytic entomopathogens on sweet sorghum and on the larvae of the stalk host, Cirsium arvense. New Phytol. 205, 816–827. borer Sesamia nonagrioides. Entomol. Exp. Appl. 154, 78–87. Hirano, E., Koike, M., Aiuchi, D., Tani, M., 2008. Pre-inoculation of cucumber roots Martinuz, A., Schouten, A., Menjivar, R.D., Sikora, R.A., 2012. Effectiveness of with Verticillium lecanii (Lecanicillium muscarium) induces resistance to systemic resistance toward Aphis gossypii (Aphididae) as induced by combined powdery mildew. Res. Bull. Obihiro Univ. 29, 82–94. applications of the endophytes Fusarium oxysporum Fo162 and Rhizobium etli Hu, G., St. Leger, R.J., 2002. Field studies using a recombinant mycoinsecticide G12. Biol. Control 62, 206–212. (Metarhizium anisopliae) reveal that it is rhizosphere competent. Appl. Environ. McKinnon, A.C., Saari, S., Moran-Diez, M.E., Meyling, N.V., Raad, M., Glare, T.R., 2017. Microbiol. 68, 6383–6387. Beauveria bassiana as an endophyte: a critical review on associated Humber, R.A., 2008. Evolution of entomopathogenicity in fungi. J. Invertebr. Pathol. methodology and biocontrol potential. Biocontrol 62, 1–17. 98, 262–266. Mutune, B., Ekesi, S., Niassy, S., Matiru, V., Bii, C., Maniania, N.K., 2016. Fungal Hyde, K.D., Soytong, K., 2008. The fungal endophyte dilemma. Fungal Divers 33, endophytes as promising tools for the management of bean stem maggot 163–173. Ophiomyia phaseoli on beans Phaseolus vulgaris. J. Pest. Sci. 89, 993–1001. Inbar, J., Menendez, A., Chet, I., 1996. Hyphal interaction between Trichoderma Muvea, A.M., Meyhofer, R., Subramanian, S., Poehling, H.-M., Ekesi, S., Maniania, N. harzianum and Sclerotinia sclerotiorum and its role in biological control. Soil Biol. K., 2014. Colonization of onions by endophytic fungi and their impacts on the Biochem. 28, 757–763. biology of Thrips tabaci. PLoS One 9 (9), e108242. Inglis, G.D., Goettel, M.S., Butt, T.M., Strasser, H., 2001. Use of hyphomycetous fungi Nahar, P.B., Kulkarni, S.A., Kulye, M.S., Chavan, S.B., Kulkarni, G., Rajendran, A., for managing insect pests. In: Butt, T.M., Jackson, C., Magan, N. (Eds.), Fungi as Yadav, P.D., Shouche, Y., Deshpande, M.V., 2008. Effect of repeated in vitro sub Biocontrol Agents. Progress, Problems and Potential. CABI Publishing, culturing on the virulence of Metarhizium anisopliae against Helicoverpa Oxfordshire, pp. 23–69. armigera (Lepidoptera: Noctuidae). Biocontrol Sci. Technol. 18, 337–355. Jaber, L.R., 2015. Grapevine leaf tissue colonization by the fungal entomopathogen Ownley, B., Gwinn, K.D., Vega, F.E., 2010. Endophytic fungal entomopathogens with Beauveria bassiana s.l. and its effect against downy mildew. Biocontrol 60, 103– activity against plant pathogens: ecology and evolution. Biocontrol 55, 113– 112. 128. Jaber, L.R., Enkerli, J., 2016. Effect of seed treatment duration on growth and Ownley, B.H., Griffin, M.R., Klingeman, W.E., Gwinn, K.D., Moulton, J.K., Pereira, R.M., colonization of Vicia faba by endophytic Beauveria bassiana and Metarhizium 2008. Beauveria bassiana: endophytic colonization and plant disease control. J. brunneum. Biol. Control 103, 187–195. Invertebr. Pathol. 3, 267–270. Jaber, L.R., Enkerli, J., 2017. Fungal entomopathogens as endophytes: can they Ownley, B.H., Pereira, R.M., Klingeman, W.E., Quigley, N.B., Leckie, B.M., 2004. promote plant growth? Biocontrol Sci.Technol. 27, 28–41. Beauveria bassiana, a dual purpose biocontrol organism, with activity against Jaber, L.R., Salem, N.M., 2014. Endophytic colonisation of squash by the fungal insect pests and plant pathogens. In: Lartey, R.T., Caesar, A. (Eds.), Emerging entomopathogen Beauveria bassiana (Ascomycota: Hypocreales) for managing Concepts in Plant Health Management. Research Signpost, Kerala, pp. 256–269. Zucchini yellow mosaic virus in cucurbits. Biocontrol Sci. Technol. 24, 1096– Parsa, S., Ortiz, V., Gómez-Jiménez, M.I., Kramer, M., Vega, F.E., 2016. Root 1109. environment is a key determinant of fungal entomopathogen endophytism Jaronski, S.T., 2010. Ecological factors in the inundative use of fungal following seed treatment in the common bean, Phaseolus vulgaris. Biol. Control. entomopathogens. Biocontrol 55, 159–185. http://dx.doi.org/10.1016/j.biocontrol.2016.09.001. Jeffries, P., 1995. Biology and ecology of mycoparasitism. Can. J. Bot. 73, 1284–1290. Pava-Ripoll, M., Angelini, C., Fang, W., Wang, S., Posada, F.J., St Leger, R., 2011. The Jirakkakul, J., Cheevadhanarak, S., Punya, J., Chutrakul, C., Senachak, J., Buajarern, T., rhizosphere-competent entomopathogen Metarhizium anisopliae expresses a Tanticharoen, M., Amnuaykanjanasin, A., 2015. Tenellin acts as an iron chelator specific subset of genes in plant root exudate. Microbiology 157, 47–55. to prevent iron-generated reactive oxygen species toxicity in the Peña-Peña, A.J., Santillán-Galicia, M.T., Hernández-López, J., Guzmán-Franco, A.W., entomopathogenic fungus Beauveria bassiana. FEMS Microbiol. Lett. 362, 1–8. 2015. Metarhizium pingshaense applied as a seed treatment induces fungal Jones, K.D., 1994. Aspects of the Biology and Biological Control of the European corn infection in larvae of the white grub Anomala cincta. J. Invertebr. Pathol. 130, 9– Borer in North Carolina (Ph.D. thesis). Department of Entomology, North 12. Carolina State University. Petrini, O., 1991. Fungal endophytes in tree leaves. In: Andrews, J.H., Hirano, S.S. Kabaluk, J.T., Ericsson, J.D., 2007. Seed treatment increases yield of field corn when (Eds.), Microbial Ecology of Leaves. Springer-Verlag, New York, pp. 179–197. applied for wireworm control. Agron. J. 99, 1377–1381. Pieterse, C.M., Zamioudis, C., Berendsen, R.L., Weller, D.M., Van Wees, S.C., Bakker, P. Karthiba, L., Saveetha, K., Suresh, S., Raguchander, T., Saravanakumar, D., A., 2014. Induced systemic resistance by beneficial microbes. Annu. Rev. Samiyappan, D., 2009. PGPR and entomopathogenic fungus bioformulation for Phytopathol. 52, 347–375. the synchronous management of leaffolder pest and sheath blight disease of Posada, F., Aime, M.C., Peterson, S.W., Rehner, S.A., Vega, F.E., 2007. Inoculation of rice. Pest Manag. Sci. 66, 555–564. coffee plants with the fungal entomopathogen Beauveria bassiana (Ascomycota: Khan, A.L., Hamayun, M., Khan, S.A., Kang, S.M., Shinwari, Z.K., Kamran, M., Ur Hypocreales). Mycol. Res. 111, 749–758. Rehman, S., Kim, J.G., Lee, I.J., 2012. Pure culture of Metarhizium anisopliae LHL07 Posada, F., Vega, F.E., 2005. Establishment of the fungal entomopathogen Beauveria reprograms soybean to higher growth and mitigates salt stress. World J. bassiana (Ascomycota: Hypocreales) as an endophyte in cocoa seedlings Microbiol. Biotechnol. 28, 1483–1494. (Theobroma cacao). Mycologia 97, 1195–1200. Krasnoff, S., Keresztes, I., Donzelli, B., Gibson, D., 2014. Metachelins, mannosylated Powell, W.A., Klingeman, W.E., Ownley, B.H., Gwinn, K.D., Dee, M., Flanagan, P.C., and N-oxidized coprogen-type metachelins, mannosylated and N-oxidized 2007. Endophytic Beauveria bassiana in tomatoes yields mycosis in tomato coprogen-type. J. Nat. Prod. 77, 1685–1692. fruitworm larvae. HortScience 42, 933.

Quesada-Moraga, E., Landa, B.B., Muñoz-Ledesma, J., Jiménez-Díaz, R.M., Santiago- rhizobacteria, Pseudomonas fluorescens for simultaneous management of Álvarez, C., 2006. Endophytic colonization of opium poppy, Papaver somniferum, leafminers and collar rot disease in groundnut. Biocontrol Sci. Technol. 20, by an entomopathogenic Beauveria bassiana strain. Mycopathologia 161, 323– 449–464. 329. Senthilraja, G., Anand, T., Durairaj, C., Raguchander, T., Samiyappan, R., 2010b. Quesada-Moraga, E., López-Díaz, C., Landa, B.B., 2014. The Hidden habit of the Chitin-based bioformulation of Beauveria bassiana and Pseudomonas fluorescens entomopathogenic fungus Beauveria bassiana: first demonstration of vertical for improved control of leafminer and collar rot in groundnut. Crop Prot. 29, plant transmission. PLoS One 9 (2), e89278. http://dx.doi.org/10.1371/journal. 1003–1010. pone.0089278. Shrivastava, G., Ownley, B.H., Augé, R.M., Toler, H., Dee, M., Vu, A., Köllner, T.G., Quesada-Moraga, E., Muñoz-Ledesma, F., Santiago-Alvarez, C., 2009. Systemic Chen, F., 2015. Colonization by arbuscular mycorrhizal and endophytic fungi protection of Papaver somniferum L. against Iraella luteipes (Hymenoptera: enhanced terpene production in tomato plants and their defense against a Cynipidae) by an endophytic strain of Beauveria bassiana (Ascomycota: herbivorous insect. Symbiosis 65, 65–74. Hypocreales). Environ. Entomol. 38, 723–730. Spatafora, J.W., Sung, G.-H., Sung, J.-M., Hywel-Jones, N.L., White, J.F., 2007. Reay, S.D., Brownbridge, M., Gicquel, B., Cummings, N.J., Nelson, T.L., 2010. Isolation Phylogenetic evidence for an animal pathogen origin of ergot and the grass and characterization of endophytic Beauveria spp. (Ascomycota: Hypocreales) endophytes. Mol. Ecol. 16, 1701–1711. from Pinus radiata in New Zealand forests. Biol. Control 54, 52–60. St. Leger, R.J., 2008. Studies on adaptations of Metarhizium anisopliae to life in the Resquín-Romero, G., Garrido-Jurado, Delso C., Ríos-Moreno, A., Quesada-Moraga, E., soil. J. Invertebr. Pathol. 98, 271–276. 2016. Transient endophytic colonizations of plants improve the outcome of Steyaert, J.M., Ridgway, H.J., Elad, Y., Stewart, A., 2003. Genetic basis of foliar applications of mycoinsecticides against chewing insects. J. Invertebr. mycoparasitism: a mechanism of biological control by species of Trichoderma. Pathol. 136, 23–31. NZ. J. Crop Horticult. 31, 281–291. Richter, D.L., Dixon, T.G., Smith, J.K., 2016. Revival of saprotrophic and mycorrhizal Tefera, T., Vidal, S., 2009. Effect of inoculation method and plant growth medium on basidiomycete cultures after 30 years in cold storage in sterile water. Can. J. endophytic colonization of sorghum by the entomopathogenic fungus Beauveria Microbiol. http://dx.doi.org/10.1139/cjm-2016-0272. bassiana. Biocontrol 54, 663–669. Ríos-Moreno, A., Garrido-Jurado, I., Resquín-Romero, G., Arroyo-Manzanares, N., Vega, F.E., Goettel, M.S., Blackwell, M., Chandler, D., Jackson, M.A., Keller, S., Koike, Arce, L., Quesada-Moraga, E., 2016. Destruxin A production by Metarhizium M., Maniania, N.K., Monzo’n, A., Ownley, B.H., Pell, J.K., Rangel, D.E.N., Roy, H.E., brunneum strains during transient endophytic colonization of Solanum 2009. Fungal entomopathogens: new insights on their ecology. Fungal Ecol. 2, tuberosum. Biocontrol Sci. Technol. 26, 1574–1585. 149–159. Rosenheim, J.A., Parsa, S., Forbes, A.A., Krimmel, W.A., Law, Y.H., Segoli, M., Segoli, Vega, F.E., 2008. Insect pathology and fungal endophytes. J. Invertebr. Pathol. 98, M., Sivakoff, F.S., Zaviezo, T., Gross, K., 2011. Ecoinformatics for integrated pest 277–279. management: expanding the applied insect ecologist’s tool-kit. J. Econ. Vega, F.E., Posada, F., Catherine Aime, M., Pava-Ripoll, M., Infante, F., Rehner, S.A., Entomol. 104, 331–342. 2008. Entomopathogenic fungal endophytes. Biol. Control 46, 72–82. Russo, M.L., Pelizza, S.A., Cabello, M.N., Stenglein, S.A., Scorsetti, A.C., 2015. Vidal, S., Jaber, L.R., 2015. Entomopathogenic fungi as endophytes: plant– Endophytic colonisation of tobacco, corn, wheat and soybeans by the fungal endophyte–herbivore interactions and prospects for use in biological control. entomopathogen Beauveria bassiana (Ascomycota, Hypocreales). Biocontrol Sci. Curr. Sci. 109, 46–54. Technol. 25, 475–480. Vincent, C., Goettel, M.S., Lazarovits, G. (Eds.), 2007. Biological Control: A Global Sánchez-Rodríguez, A.R., del Campillo, M.C., Quesada-Moraga, E., 2015. Beauveria Perspective. CAB International/AAFC, Wallingford, United Kingdom. bassiana: an entomopathogenic fungus alleviates Fe chlorosis symptoms in Wainwright, M., Betts, R.P., Teale, D.M., 1986. Antibiotic activity of oosporein from plants grown on calcareous substrates. Sci. Hortic. 197, 193–202. Verticillium psalliotae. Trans. Br. Mycol. Soc. 86, 168–170. Sasan, R.K., Bidochka, M.J., 2012. The insect-pathogenic fungus Metarhizium robertsii Wang, Q., Xu, L., 2012. Beauvericin, a bioactive compound produced by fungi: a (Clavicipitaceae) is also an endophyte that stimulates plant root development. short review. Molecules 17, 2367–2377. Am. J. Bot. 99, 101–107. Wraight, S.P., Inglis, G.D., Goettel, M.S., 2007. Fungi. In: Lacey, L.A., Kaya, H.K. (Eds.), Sasan, R.K., Bidochka, M.J., 2013. Antagonism of the endophytic insect pathogenic Field Manual of Techniques in Invertebrate Pathology. second ed. The fungus Metarhizium robertsii against the bean plant pathogen Fusarium solani f. Netherlands, Dordrecht, pp. 223–248. sp. phaseoli. Can. J. Plant Pathol. 35, 288–293. Widler, B., Muller, E., 1984. Untursuchungen uber endophytische Pilze von Schultz, B., Guske, S., Dammann, U., Boyle, C., 1998. Endophyte–host interactions II. Arctostaphylos uva-ursi (L.) Sprenger (Ericaceae). Bot. Helv. 94, 307–337. Defining symbiosis of the endophytehost interaction. Symbiosis 25, 213–227. Zeilinger, S., Galhaup, C., Payer, K., Woo, S.L., Mach, R.L., Fekete, C., Lorito, M., Schulz, B., Boyle, C., 2005. The endophytic continuum. Mycol. Res. 109, 661–686. Kubicek, C.P., 1999. Chitinase gene expression during mycoparasitic interaction Schulz, B., Haas, S., Junker, C., Andree, N., Schobert, M., 2015. Fungal endophytes are of Trichoderma harzianum with its host. Fungal Genet. Biol. 26, 131–140. involved in multiple balanced antagonisms. Curr. Sci. 109, 39–45. Zhou, W., Wheeler, T.A., Starr, J.L., Valencia, C.U., Sword, G.A., 2016. A fungal Senthilraja, G., Anand, T., Durairaj, C., Kennedy, J.S., Suresh, S., Raguchander, T., endophyte defensive symbiosis affects plant-nematode interactions in cotton. Samiyappan, R., 2010a. A new microbial consortia containing Plant Soil. http://dx.doi.org/10.1007/s11104-016-3147-z. entomopathogenic fungus, Beauveria bassiana and plant growth promoting